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Objective To observe the inhibitory effect of caudal vein injection of senescence marker protein 30（SMP30） on myocardial injury in septic mice and to explore its mechanism. Methods Sixty male C57BL/6 mice were randomly divided into the Con+empty vector group, LPS+empty vector group, and LPS+SMP30 group, with 20 mice in each. Mice in the Con+empty vector group and LPS+empty vector group were injected with adeno-null associated virus vector via the tail vein, and mice in the LPS+SMP30 group were injected with SMP30 adeno-associated virus vector via the tail vein. After 2 weeks of adeno-associated virus vector injection, mice in the LPS+empty vector group and LPS+SMP30 group were intraperitoneally injected with lipopolysaccharide（LPS） to establish the sepsis models, while mice in the Con+empty vector group were given intraperitoneal injection of the same volume of normal saline. Left ventricular ejection fraction（LVEF） and left ventricular fraction shortening（LVFS） were measured by echocardiography at 24 h after LPS injection.Serum and heart tissues were collected after the mice were killed, and serum levels of creatine kinase-MK（CK-MB), lactic dehydrogenase（LDH） and cTnI, and myocardial levels of malondialdehyde（MDA), superoxide dismutase（SOD） and glutathione peroxidase（GSH-Px） were detected by ELISA. TUNEL staining was used to detect cardiomyocyte apoptosis.Macrophage infiltration was detected by immunofluorescence staining. The myocardial reactive oxide species（ROS） production was detected by dihydroethidium fluorescent probe. Interleukin（IL）-1β, tumor necrosis factor（TNF）-α and IL-6mRNA levels were detected by qRT-PCR. The protein expression levels of myocardial tissues SMP30, silent information regulator 1（SIRT1), Ac-NF-κB p65, and Ac-FOXO1 were detected by Western blotting. Results Compared with the Con+empty vector group, the values of LVEF and LVFS decreased, the levels of serum CK-MB, LDH and cTnI increased, the apoptosis rate increased, the myocardial macrophage infiltration was more obvious, the mRNA levels of IL-1β, IL-6, TNF-α increased, the productions of ROS and MDA increased, the activities of SOD and GSH-Px decreased, the expression of SIRT1 down-regulated, and the expression levels of Ac-NF-κB p65 and Ac-FOXO1 increased in the LPS+empty vector group（all P＜0. 01). Compared with LPS+empty vector group, the values of LVEF and LVFS increased, the levels of serum CK-MB, LDH and cTnI decreased, the apoptosis rate decreased, the myocardial macrophage infiltration decreased, the mRNA levels of IL-1β, IL-6, TNF-α decreased, the productions of ROS and MDA decreased, the activities of SOD and GSH-Px increased, the expression of SIRT1 increased, and the expression levels of Ac-NF-κB p65 and AcFOXO1 decreased in the LPS+SMP30 group（all P＜0. 01). Conclusion The caudal vein injection of SMP30 can up-regulate SMP30 expression in mice with sepsis, inhibit myocardial inflammatory response and oxidative stress level, thereby alleviating myocardial injury, and its mechanism may be related to activating the SIRT1 signaling pathway. [ABSTRACT FROM AUTHOR]

Human senescence marker protein 30 (huSMP30) has been characterized as a multifaceted protein consisting of various enzymatic and cellular functions. It catalyzes the interconversion of L-gulonate and L-gulono-γ-lactone in the ascorbate biosynthesis pathway. Therefore, we hypothesized that it could be a potential anti-biofilm agent against pathogenic bacteria due to its lactonase activity. In order to corroborate this, the huSMP30 was recombinantly expressed, purified, and analyzed for its ability to inhibit Mycobacterium smegmatis biofilm formation, which showed a concentration-dependent inhibition as compared to the untreated control group. Further, in silico analysis was performed to redesign the huSMP30 with enhanced lactonase activity. Molecular docking analysis of the huSMP30 and lactone substrates facilitated the selection of three single amino acid substitutions (E18H, N154Q, and D204V), which were created using a PCR-based site-directed mutagenesis reaction. These mutant proteins and the wild-type huSMP30 were purified, and the effects on the enzymatic activity and biofilm formation were studied. The mutants E18H and D204V showed non-significant effects on specific lactonase activity, catalytic efficiency, and anti-biofilm property; however, the mutant N154Q showed significant improvement in the specific lactonase activity, catalytic efficiency, and inhibition in the biofilm formation. The protein stability analysis revealed that the wild-type huSMP30 and its designed mutants were stable at 37 °C for up to 4 days. In conclusion, the anti-biofilm property of the huSMP30 has been established, and an engineered version, N154Q, inhibits biofilm formation with greater efficiency. Human SMP30 is a versatile protein with multiple cellular and enzymatic functions, however, its anti-biofilm potential has not been explored. Our work presents the method to produce soluble and active huSMP30 in the E. coli expression system and establishes its role as an anti-biofilm agent against Mycobacterium smegmatis owing to its lactonase activity. Our results provide support for the future advancement of huSMP30 as a potential anti-biofilm agent targeting pathogenic Mycobacterium species. [ABSTRACT FROM AUTHOR]

Background/Aim: Chronic kidney disease (CKD) is one of the most common causes of mortality in wild nondomestic felidae. The molecular mechanism regulating renal fibrosis in nephropathy is not fully understood especially in the felidae. This study aimed to elucidate senescence marker protein 30 (SMP30) expression patterns and its relationship with epithelial-mesenchymal transition (EMT) by immunostaining in two necropsied Siberian tigers (Panthera tigris altaica) with CKD. Materials and Methods: Two kidney samples from male Siberian tigers were fixed and tissue sections were stained for histopathological assay. Results: In CKD, renal tubular epithelial cells lost their tubular structures surrounded by severe interstitial fibrosis and were detached from the basement membrane. These damaged cells resembled the morphology of mesenchymal cells and showed much lower SMP30 expression compared with intact tubular epithelial cells. These cells also expressed vimentin, which is specifically expressed by mesenchymal cells, and through double staining, it was observed that vimentin was expressed in the tubular epithelial cells where SMP30 was not expressed. In addition, double-positive expression of pan-cytokeratin (pan-CK) and vimentin was found in damaged epithelial cells with mesenchymal features. Conclusion: We demonstrated possible evidence to understand the role of SMP30 as a new pivotal factor and the possibility of decreased SMP30 as a potential indicator of EMT at the end stage of CKD. [ABSTRACT FROM AUTHOR]

Background: The senescence marker protein 30 (SMP30) is a calcium-binding protein whose expression decreases with age, and is closely associated with hepatocellular carcinoma (HCC) development. The primary goal of this study was to examine the mechanistic effect of SMP30 on HCC migration and invasion. Methods: Bioinformatic and immunohistochemical approaches were used to examine the expression of SMP30 in HCC tissues and its relationship to patient survival. We investigated the effects of SMP30 expression on HCC cell proliferation, migration, invasion, and cell cycle dynamics. cDNA microarray technology was used to determine the gene expression profile of SK-Hep-1 cells following recombinant SMP30 overexpression to identify genes downstream of SMP30 that regulate HCC cell migration and invasion. We identified SMP30 interacting proteins by affinity purification-mass spectrometry (AP-MS) and co-immunoprecipitation/western blotting (COIP-WB). Results: SMP30 expression was lower in HCC tissues compared with normal liver tissues, and its expression positively correlated with overall survival in HCC patients. Additionally, SMP30 overexpression effectively blocked the migratory and invasive properties of SK-Hep-1 cells, but did not affect either proliferation rates or cell cycle. cDNA microarray results confirmed that many of the differentially expressed genes identified are involved in the process of epithelial-mesenchymal transition (EMT). AP-MS and COIP-WB experiments confirmed that Rho-associated protein kinase 1 (ROCK1) interacts with SMP30 in SK-Hep-1 cells, and ROCK1 is known to intimately regulate the EMT process. Conclusion: SMP30 inhibits HCC metastasis by influencing the expression of EMT-related proteins after interacting with ROCK1.

AIM: To explore the effect of low expression of senescence marker protein 30(SMP30)on proliferation and oxidative stress of human lens epithelial cells(LECs)line SRA01/04 under high calcium conditions. METHODS: Three RNAi sequences were designed to knock down SMP30 target gene RGN expression(KD1-3), and the blank-load sequence was used as the negative control group(NCKD), all of which were used to construct lentiviral vectors to infect SRA01/04 cells. Meanwhile, the uninfected SRA01/04 cells was used as the blank control group(CON). After transfecting SRA01/04 cells, the lentiviral vector with the highest knockdown efficiency was selected by RT-PCR for subsequent experiments. Cells were treated with 15mmol/L CaCl2 for 24h to simulate a high calcium conditions. BrdU-Elisa assay was used to measure cell proliferation, superoxide dismutase(SOD)assay kit and oxidized glutathione/total glutathione(GSSG/T-GSH)assay kit were used to detect the level of intracellular oxidative stress. RESULTS: KD1-3 and NCKD lentiviral vectors were successfully constructed to infect SRA01/04 cells with an infection efficiency of about 80%. The knockdown efficiency of KD1-3 group was 93%, 60% and 74%, respectively, KD1 group was selected for follow-up experiment. Under the high calcium conditions, the activity of relative cell proliferation and SOD in KD1 group \〖(2.42±0.08)and(11.69±0.52U/mg)\〗 were significantly lower than that in NCKD group \〖(2.95±0.08)and(31.10±2.24U/mg)\〗 and CON group \〖(2.96±0.25)and(26.33±1.04U/mg)\〗, the ratio of GSSG/T-GSH in KD1 group(70.80±2.34)was significantly higher than that in NCKD group(15.93±3.47)and CON group(20.05±2.45)(PP>0.05).CONCLUSION: Under high calcium conditions, SRA01/04 cells(HLECs)with low expression of SMP30 mediated by shRNA lentivirus resulted in the decrease of the proliferation activity and antioxidant capacity, suggesting that SMP30 may play a protective role in regulating cell proliferation and anti-oxidative stress in HLECs.

Background: The senescence marker protein 30 (SMP30) is a calcium-binding protein whose expression decreases with age, and is closely associated with hepatocellular carcinoma (HCC) development. The primary goal of this study was to examine the mechanistic effect of SMP30 on HCC migration and invasion., Methods: Bioinformatic and immunohistochemical approaches were used to examine the expression of SMP30 in HCC tissues and its relationship to patient survival. We investigated the effects of SMP30 expression on HCC cell proliferation, migration, invasion, and cell cycle dynamics. cDNA microarray technology was used to determine the gene expression profile of SK-Hep-1 cells following recombinant SMP30 overexpression to identify genes downstream of SMP30 that regulate HCC cell migration and invasion. We identified SMP30 interacting proteins by affinity purification-mass spectrometry (AP-MS) and co-immunoprecipitation/western blotting (COIP-WB)., Results: SMP30 expression was lower in HCC tissues compared with normal liver tissues, and its expression positively correlated with overall survival in HCC patients. Additionally, SMP30 overexpression effectively blocked the migratory and invasive properties of SK-Hep-1 cells, but did not affect either proliferation rates or cell cycle. cDNA microarray results confirmed that many of the differentially expressed genes identified are involved in the process of epithelial-mesenchymal transition (EMT). AP-MS and COIP-WB experiments confirmed that Rho-associated protein kinase 1 (ROCK1) interacts with SMP30 in SK-Hep-1 cells, and ROCK1 is known to intimately regulate the EMT process., Conclusion: SMP30 inhibits HCC metastasis by influencing the expression of EMT-related proteins after interacting with ROCK1., Competing Interests: The authors declare no conflict of interest., (© 2024 The Author(s). Published by IMR Press.)

Sitagliptin (SIT) is an antidiabetic used worldwide to ameliorate the hyperglycemia and insulin insensitivity induced dysmetabolism. In this study, we investigated the effect of sitagliptin and vitamin E on metabolic dysfunction in high-fat diet (HFD) fed rats. Sixty-four male rats were allocated into 8 groups (n = 8) as follow; control, control + vitamin E, control + sitagliptin, control + sitagliptin + vitamin E, HFD, HFD + vitamin E, HFD + sitagliptin and HFD + sitagliptin + vitamin E. Control groups were fed with chow diet for 15 weeks, while HFD groups were fed with HFD for the same duration. Vitamin E and sitagliptin were administered in the last 4 weeks of the study. At the end of the 15th week, body weight, liver weight/body weight ratio, weight gain, glucose, lipid profile, liver enzymes, adiponectin and proinflammatory cytokines as interleukin 6 (IL-6), high sensitive C reactive protein (hs-CRP) and tumour necrosis factor-α (TNF-α) were measured. Additionally, gene expressions of senescence marker protein 30 (SMP30), Bcl-2, and Bax were measured. Total antioxidant capacity (TAC) and thiobaribituric acid reactive substances (TBARS) were assayed. HFD increased TBARS, IL-6, hs-CRP and TNF-α significantly and decreased TAC and adiponectin. Sitagliptin produced a comparable result through increasing adiponectin, sitagliptin alone or in combination with vitamin E increased the TAC, and gene expression of SMP30 and Bcl-2 and decreased TBARS with downregulation of the overexpressed Bax. Vitamin E, as a natural antioxidant, ameliorates the oxidative stress with insignificant change in lipid profile and inflammatory cytokine levels. Concomitant sitagliptin and vitamin E reduced the hepatic dysfunction induced by HFD. [ABSTRACT FROM AUTHOR]

Background/aim: Chronic kidney disease (CKD) is one of the most common causes of mortality in wild non-domestic felidae. The molecular mechanism regulating renal fibrosis in nephropathy is not fully understood especially in the felidae. This study aimed to elucidate senescence marker protein 30 (SMP30) expression patterns and its relationship with epithelial-mesenchymal transition (EMT) by immunostaining in two necropsied Siberian tigers (Panthera tigris altaica) with CKD., Materials and Methods: Two kidney samples from male Siberian tigers were fixed and tissue sections were stained for histopathological assay., Results: In CKD, renal tubular epithelial cells lost their tubular structures surrounded by severe interstitial fibrosis and were detached from the basement membrane. These damaged cells resembled the morphology of mesenchymal cells and showed much lower SMP30 expression compared with intact tubular epithelial cells. These cells also expressed vimentin, which is specifically expressed by mesenchymal cells, and through double staining, it was observed that vimentin was expressed in the tubular epithelial cells where SMP30 was not expressed. In addition, double-positive expression of pan-cytokeratin (pan-CK) and vimentin was found in damaged epithelial cells with mesenchymal features., Conclusion: We demonstrated possible evidence to understand the role of SMP30 as a new pivotal factor and the possibility of decreased SMP30 as a potential indicator of EMT at the end stage of CKD., (Copyright © 2024, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Human senescence marker protein 30 (huSMP30) has been characterized as a multifaceted protein consisting of various enzymatic and cellular functions. It catalyzes the interconversion of L-gulonate and L-gulono-γ-lactone in the ascorbate biosynthesis pathway. Therefore, we hypothesized that it could be a potential anti-biofilm agent against pathogenic bacteria due to its lactonase activity. In order to corroborate this, the huSMP30 was recombinantly expressed, purified, and analyzed for its ability to inhibit Mycobacterium smegmatis biofilm formation, which showed a concentration-dependent inhibition as compared to the untreated control group. Further, in silico analysis was performed to redesign the huSMP30 with enhanced lactonase activity. Molecular docking analysis of the huSMP30 and lactone substrates facilitated the selection of three single amino acid substitutions (E18H, N154Q, and D204V), which were created using a PCR-based site-directed mutagenesis reaction. These mutant proteins and the wild-type huSMP30 were purified, and the effects on the enzymatic activity and biofilm formation were studied. The mutants E18H and D204V showed non-significant effects on specific lactonase activity, catalytic efficiency, and anti-biofilm property; however, the mutant N154Q showed significant improvement in the specific lactonase activity, catalytic efficiency, and inhibition in the biofilm formation. The protein stability analysis revealed that the wild-type huSMP30 and its designed mutants were stable at 37 °C for up to 4 days. In conclusion, the anti-biofilm property of the huSMP30 has been established, and an engineered version, N154Q, inhibits biofilm formation with greater efficiency. Human SMP30 is a versatile protein with multiple cellular and enzymatic functions, however, its anti-biofilm potential has not been explored. Our work presents the method to produce soluble and active huSMP30 in the E. coli expression system and establishes its role as an anti-biofilm agent against Mycobacterium smegmatis owing to its lactonase activity. Our results provide support for the future advancement of huSMP30 as a potential anti-biofilm agent targeting pathogenic Mycobacterium species., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Objective To investigate the effects of human hepatoma-associated antigen senescence marker protein 30 (SMP30) on the maturation of human peripheral blood dendritic cells (DCs). Methods Peripheral blood mononuclear cells were isolated from healthy adult peripheral blood by Ficoll density gradient centrifugation. DCs were induced by granulocyte-macrophage colony-stimulating factor and interleukin-4.We identified the DCs by flow cytometry. DCs were randomly divided into the SMP30 lentivirus group and empty vector group, which were transfected with human hepatoma-associated antigen SMP30 recombinant lentivirus and empty lentivirus, respectively; and a blank control group was also set up. The expression of green fluorescent protein in each group was observed under fluorescence microscope to determine the transfection of DCs. The expression of SMP30 in DCs of each group was detected by Western blotting. The secretion of IL-12 and INF-γ in DCs of each group was detected by ELISA. Flow cytometry was used to detect the expression of co-stimulatory molecules CD80 and CD86 on DCs surface. Results Green fluorescent protein was successfully expressed in DCs of the SMP30 lentivirus group and empty lentivirus group under fluorescence microscope. However, the expression of green fluorescent protein was not observed in DCs of the blank control group. Compared with the empty lentivirus group and the blank control group, the SMP30 lentivirus group had higher expression of SMP30, higher secretion of IL-12 and INF-γ, and higher expression rates of CD80 and CD86 (all P < 0.05). Conclusion In vitro transfection of human hepatoma-associated antigen SMP30 lentivirus can promote the maturation of human peripheral blood DCs. [ABSTRACT FROM AUTHOR]

Organophosphorus compounds (OPCs) are able to interact with various biological targets in living organisms, including enzymes. The binding of OPCs to enzymes does not always lead to negative consequences for the body itself, since there are a lot of natural biocatalysts that can catalyze the chemical transformations of the OPCs via hydrolysis or oxidation/reduction and thereby provide their detoxification. Some of these enzymes, their structural differences and identity, mechanisms, and specificity of catalytic action are discussed in this work, including results of computational modeling. Phylogenetic analysis of these diverse enzymes was specially realized for this review to emphasize a great area for future development(s) and applications.

To our knowledge, this is the first study to report down-regulation of senescence marker protein 30 (SMP30) by iron-specific chelator deferoxamine (DFO) on FAO cell senescence, using a DNA microarray. Furthermore, DFO treatment increased senescence marker β-galactosidase activity, whereas this activity was attenuated by overexpression of SMP30. Our data suggested that down-regulation of SMP30 drives cell senescence in iron-chelated condition. [ABSTRACT FROM AUTHOR]

Senescence marker protein 30 (SMP30) has been reported to serve antiapoptotic and antioxidant roles, as well as roles in Ca2+ regulation, and may be involved in the occurrence and development of cataract. The present study aimed to investigate the expression of SMP30 in senescent human lens epithelial cells (HLECs) and explored the relationship between SMP30 and aging. SRA01/04 cells, a HLEC line, were treated with H2O2 to mimic aging, and cell morphological changes were observed by microscopy and cell activity was examined by MTT assay, senescence‑associated‑β‑galactosidase (SA‑β‑Gal) staining and cell cycle analysis. The expression of SMP30 mRNA and protein was measured by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blotting. Following prolonged low‑dose H2O2 exposure, cells exhibited senescence‑related morphological changes, reduced growth activity, increased SA‑β‑Gal positive staining and cell cycle arrest in the S and G2/M phases. SMP30 mRNA expression levels were significantly downregulated following exposure to 75 and 100 μM H2O2, and the protein expression levels in the same groups were decreased by >6‑fold compared with the control untreated cells. However, no significant change was observed in SMP30 expression in the 25 and 50 μM H2O2 exposure groups. These results suggest that, in the early stage of senescence induced by H2O2‑mediated chronic oxidative stress, there may be no significant change in SMP30 expression, but when the oxidative stress increases and senescence is aggravated, SMP30 may be significantly downregulated in the senescent HLECs. The present study indicates that SMP30 may be an important factor involved in the aging process of HLECs and the development of cataract. [ABSTRACT FROM AUTHOR]

Objective To determine appropriate CpG sites for senescence marker protein 30 (SMP30) promoter methyla-tion detection in hepatocellular carcinoma using pyrosequencing. Methods Genomic DNA was isolated from human hepatocel-lular carcinoma cell lines SMCC-7721 and BEL-7404. The amplification and sequencing primers were designed for SMP30 gene sequence 1 and sequence 2, and the methylation status of CpG sites in SMP30 gene sequence 1 and sequence 2 was determined by pyrosequencing technology. Sanger sequencing was used to validate the SNP. Results In SMCC-7721 and BEL-7404 cell lines, the 6 CpG sites in SMP30 gene sequence 1 were low methylatied. which was not consistent with die low expression of SMP30 gene, and the quality of the sequencing results all failed. The 6 CpG sites in the SMP30 gene sequence 2 were highly mediylated. which was consistent with the low expression of SMP30 gene, the first 2 CpG sites passed, and the last 4 CpG sites failed. The 5th and 6th CpG sites were ignored and only die first 4 CpG sites before the poly (T) structure in SMP30 gene sequence 2 were detected, the percentage of methylation in BCC-7721 cell line was 100% . 90% . 100% and 80%. The percentage of methylation in BEL-7404 cell line was 100% . 92% . 100% and 77%. Except for the 3th CpG site, the rest of the CpG sites passed. Sanger sequencing showed diat SMCC-7721 and BEL-7404 cell lines had SNP sites before the 3di CpG site, and all of them were A. Conclusion Pyrosequencing methylation analysis of CpG sites near die upstream of transcription start site and he-fore the poly (T) structure in SMP30 promoter is more accurate. [ABSTRACT FROM AUTHOR]

Objectives: Tumor immunotherapy based on dendritic cells (DC) is one of the most promising approaches to treat cancers. This therapy uses an immunogenic tumor antigen to present it to T cells. Senescence marker protein 30 (SMP30) is identified as a tumor associated antigen (TAA) with high immunogenicity and specificity for hepatocellular carcinoma (HCC). DCs are the most potent antigen presenting cells, and can be transduced with tumor antigens to enhance antitumor immune response. The purpose of this study was to investigate the antitumor effect of DCs transduced with a recombinant lentiviral vector ( LV-SMP30 ) expressing SMP30., Methods: A recombinant lentiviral vector ( LV-SMP30 ) expressing SMP30 was constructed and transduced into DCs. The expression of SMP30 was detected by western blot. Mouse bone marrow-derived DCs were divided into four groups: LV-SMP30 group (transduced with LV-SMP30 ), Protein group (co-cultured with SMP30 protein), LV group (transduced with the empty vector) and Untreated group (the normal DCs). The effect of LV-SMP30 on DCs was detected through surface markers (CD123, CD11c, CD80 and CD86) and cytokine production. The activation and proliferation of CD3 + CD8 + T cells were detected by CCK-8 kit. Flow cytometry was used to detect CD3 + CD8 + T cell-mediated cytotoxicity. After construction of a mouse subcutaneous xenograft model, the volume and growth of tumors in different groups were observed. The changes in serum immune indexes in the treated groups were compared with those in the control group., Results: The LV-SMP30 recombinant was constructed and transduced into DCs successfully, and LV-SMP30 -transduced DCs stably expressed SMP30. The percentages of expression in the LV-SMP30 and Protein groups were significantly higher than those in the LV or Untreated groups ( P<0.05 ). Meanwhile, after the DCs were cultured for 72 hours, the levels of IL-2, IL-6, IL-12, and IFN-γ were significantly higher in the LV-SMP30 and Protein groups than in the LV group or Untreated group ( P <0.05). After the DCs were continuously cultured for one week, however, the cytokine levels in the LV-SMP30 group were significantly higher than those in the Protein group ( P <0.05). In addition, CD3 + CD8 + T cell proliferation and activation levels were substantially higher in the LV-SMP30 and Protein groups than in the LV or Untreated groups ( P <0.05). Furthermore, as the ratio of effectors/target cells increasing in the LV-SMP30 group, CD3 + CD8 + T cell-mediated cytotoxicity in H22 cells became higher (0:1, 10:1; 20:1; 40:1, respectively). In comparison to the control group, the cytotoxicity of the LV-SMP30 group was considerably increased at the ratios of 10:1, 20:1 and 40:1 ( P <0.05). However, in the case of Hep1-6 cells, there was no significant difference in CD3 + CD8 + T cell-mediated cytotoxicity among the groups. In addition, when compared with other groups, the mice in the LV-SMP30 group showed the most volume reduction, the slowest tumor growth, and the highest level of IL-2 and IFN-γ ( P <0.05)., Conclusion: DCs transduced with LV-SMP30 can dramatically enhance specific CD3 + CD8 + T cell immune responses against mouse hepatocarcinoma cells in vitro and in vivo. These findings lend significant support to the development of the DC-based SMP30 antigen vaccine for hepatocarcinoma immunotherapy., Competing Interests: None., (AJTR Copyright © 2022.)

Senescence marker protein-30 (SMP30) has been identified as the lactone-hydrolysing enzyme gluconolactonase (GNL), which is involved in vitamin C (l-ascorbic acid, AA) biosynthesis. We previously reported the development of SMP30/GNL knockout (KO) mice unable to synthesize AA in vivo. For more efficient study of the liver's AA uptake and as yet uncharacterized efflux system, we established an immortal hepatocyte line derived from a hybrid of SMP30/GNL KO mice and Immortomice. Immortomice express the thermolabile simian virus 40 (SV40) large T antigen tsA58. These SMP30/GNL KO immortal hepatocytes proliferate at the permissive temperature of 33°C but degrade rapidly at the non-permissive temperature of 39°C. Additionally, they are SMP30-/GNL-deficient, express SV40 large T antigen and proliferate steadily at 33°C. However, the cells’ proliferation is arrested at 39°C. A phase contrast micrograph revealed that the cells are binucleated with an enlarged cytoplasm similar to that of primary cultured hepatocytes from wild-type mice. Dose–response and time-dependent study of AA uptake revealed that the cells, although unable to synthesize AA, took up AA from the culture medium. This property of our SMP30/GNL immortal hepatocytes makes them extremely useful for studying AA uptake and efflux systems in the liver. [ABSTRACT FROM PUBLISHER]

Abstract: Using senescence marker protein 30 (SMP30)/gluconolactonase (GNL) knockout (KO) mice, which cannot synthesize vitamin C (VC), we examined whether modulating VC level affects age-related hearing loss (AHL). KO and wild-type (WT) C57BL/6 mice were given water containing 1.5g/L VC [VC(+)] or 37.5mg/L VC [VC(−)]. At 10months of age, KO VC(−) mice showed significant reduction in VC level in the inner ear, plasma, and liver, increase in auditory brainstem response (ABR) thresholds, and decrease in the number of spiral ganglion cells compared to WT VC(−), WT VC(+), and KO VC(+) mice. There were no differences in VC level in the inner ear, ABR thresholds, or the number of spiral ganglion cells among WT VC(−), WT VC(+), and KO VC(+) mice. These findings suggest that VC depletion can accelerate AHL but that supplementing VC may not increase VC level in the inner ear or slow AHL in mice. [Copyright &y& Elsevier]

Senescence marker protein 30 (SMP30) is identified as an important aging marker molecule and known to play multifunctional roles as an intracellular calcium regulatory protein in the signaling process. To elucidate the functional significance of SMP30, we established the stably transfected P19 cell line with SMP30 expression vector. Overexpression of SMP30 slightly suppressed the proliferation of P19 cells. However, SMP30 overexpression was cytoprotective against calcium-mediated stress such as calcium ionophore (A23187), and thapsigargin. We found that SMP30 overexpression reduced the elevated intracellular calcium levels induced by A23187, but not by thapsigargin. In addition, SMP30 transfected P19 cells were more protective to tert-butylhydroperoxide induced cytotoxicity, indicating the antioxidative properties of SMP30. Taken together, our results suggest that external calcium regulation and antioxidant properties are involved in the cytoprotective mechanism of SMP30. [ABSTRACT FROM AUTHOR]

Abstract: There are large inter-individual differences in the metabolism of vitamin C (VC), which is composed of both ascorbic acid (AsA) and dehydroascorbic acid (DAsA). AsA is oxidized to DAsA in a series of xenobiotic reactions. Thus, the effects of polymorphism A313G (Ile105Val) in the gene for glutathione S-transferases P1 (GSTP1), one of the most active xenobiotic enzymes, on human VC metabolism were studied. The variant frequency of GSTP1 among the present subjects (n =210) was AA 71.0%; GA 27.0% and GG 1.9%. At 24h after administration of 1mmol of VC to young women (n =17; age, 21.0±1.1 y), total VC excretion (46.7±18.1mg) by AA homozygotes of GSTP1 was greater (p <0.0069) than that (28.2±14.0mg) by GA heterozygotes. One hour after administration of VC, blood total VC levels were also significantly different (p <0.0036) between the homozygotes and heterozygotes. The effects of other polymorphisms in xenobiotic enzymes on VC metabolism were small. [Copyright &y& Elsevier]

Etiology and pathogenesis of age-related cataract is not entirely clear till now. Senescence marker protein 30 (SMP30) is a newly discovered anti-aging factor, which plays an important role in preventing apoptosis and reducing oxidative stress damage. Mitochondria are located at the intersection of key cellular pathways, such as energy substrate metabolism, reactive oxygen species (ROS) production and apoptosis. Oxidative stress induced by 4-hydroxynonenal (4-HNE) is closely related to neurodegenerative diseases and aging. Our study focused on the effect of SMP30 on mitochondrial homeostasis of human lens epithelial cells (HLECs) induced by 4-HNE. Western blots and qPCR were used to compare the expression of SMP30 protein in the residual lens epithelial cells in the lens capsule of age-related cataract (ARC) patients and the donated transparent lens capsule. On this basis, SMP30 overexpression plasmid and SMP30 shRNA interference plasmid were introduced to explore the effect of SMP30 on the biological behavior in HLECs under the condition of oxidative stress induced by 4-HNE through immunohistochemistry, ROS evaluation, metabolic spectrum analysis and JC-1 fluorescence measurement. Given that Nuclear Factor erythroid 2-Related Factor 2 (Nrf2)/Kelch Like ECH Associated Protein 1 (KEAP1) signaling pathway is the most important antioxidant stress pathway, we further analyzed the regulatory effect of SMP30 by WB to explore its molecular mechanism. Our study indicated that SMP30 may inhibit ROS accumulation, restore mitochondrial function, activate Nrf2/Keap1 signaling pathway, therefore protecting lens epithelial cells from oxidative stress-induced cell damage.

Organophosphorus compounds (OPCs) are able to interact with various biological targets in living organisms, including enzymes. The binding of OPCs to enzymes does not always lead to negative consequences for the body itself, since there are a lot of natural biocatalysts that can catalyze the chemical transformations of the OPCs via hydrolysis or oxidation/reduction and thereby provide their detoxification. Some of these enzymes, their structural differences and identity, mechanisms, and specificity of catalytic action are discussed in this work, including results of computational modeling. Phylogenetic analysis of these diverse enzymes was specially realized for this review to emphasize a great area for future development(s) and applications. [ABSTRACT FROM AUTHOR]

Aim: To study the effect of senescence marker protein 30 (SMP30) on the proliferation and apoptosis of human lens epithelial cell (HLEC) SRA01/04., Methods: SMP30 overexpression (OE) and knock down (KD) type cell lines were cultivated by using two groups regucalcin (RGN; SMP30) lentiviral vectors (LV-RGN, LV-RGN-RNAi) and the respective negative control virus infect SRA01/04 cells. Western blot and real-time quantitative polymerase chain reaction (q-PCR) analysis were used to determine RGN overexpression and knock down efficiency. We use cell counting kit-8 (CCK8) assay to measure cell viability and 5-bromodeoxyuridine (BrdU) assay to test cell proliferation. Cell cycle was measured by PI FACS assay and cell apoptosis was tested by Annexin V-APC assay through flow cytometry. We use Western blot to measure the content of caspase-3 in SRA01/04., Results: We used PCR and Western blot techniques to determine the successful transfection of SMP30 OE and KD SRA01/04 cell lines. By CCK8, Brdu and PI FACS cell cycle assay, it was found that the SMP30 OE group promoted cell proliferation ( P <0.05) compared with the control group, and the KD group inhibited cell proliferation ( P <0.05). The results of Annexin V-APC signal staining detection indicated that compared with respective control group, the cell apoptosis rate was higher in KD group ( P <0.05) but lower in OE group ( P <0.01). The expression of caspase-3 was down-regulated in OE group through Western blot assay and up-regulated in KD group compared with respective control group., Conclusion: Proliferation of SRA01/04 was promoted by SMP30 OE and apoptosis was suppressed. Increasing the expression of SMP30 may protect HLEC SRA01/04 against apoptosis in cataract.

Background and Objective: Senescence marker protein 30 (SMP30) is assumed to behave as an anti-aging factor. Recently, we have demonstrated that deficiency of SMP30 exacerbates angiotensin II-induced cardiac hypertrophy, dysfunction and remodeling, suggesting that SMP30 may have a protective role in the heart. Thus, this study aimed to test the hypothesis that up-regulation of SMP30 inhibits cardiac adverse remodeling in response to angiotensin II., Methods: We generated transgenic mice with cardiac-specific overexpression of SMP30 gene using α-myosin heavy chain promoter. Transgenic mice and wild-type littermate mice were subjected to continuous angiotensin II infusion (800 ng/kg/min)., Results: After 14 days, heart weight and left ventricular weight were lower in transgenic mice than in wild-type mice, although blood pressure was similarly elevated during angiotensin II infusion. Cardiac hypertrophy and diastolic dysfunction in response to angiotensin II were prevented in transgenic mice compared with wild-type mice. The degree of cardiac fibrosis by angiotensin II was lower in transgenic mice than in wild-type mice. Angiotensin II-induced generation of superoxide and subsequent cellular senescence were attenuated in transgenic mouse hearts compared with wild-type mice., Conclusions: Cardiac-specific overexpression of SMP30 inhibited angiotensin II-induced cardiac adverse remodeling. SMP30 has a cardio-protective role with anti-oxidative and anti-aging effects and could be a novel therapeutic target to prevent cardiac hypertrophy and remodeling due to hypertension., (Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.)